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. 2015 Mar 15;26(6):1141–1159. doi: 10.1091/mbc.E14-07-1222

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© 2015 Garcia-Alvarez et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 3: — STIM2 shapes functional synaptic inputs. (A–D) whole-cell patch-clamp recordings of hippocampal neurons (DIV 15–18) sparsely transfected with the indicated shRNA constructs. (A) Individual mEPSC traces of cells expressing the indicated shRNAs. STIM2 shRNA#1 was coexpressed with YFP-STIM2 for rescue experiments. (B–D) Quantification of mEPSC frequency and amplitude (Scr, 23.2 ± 2 pA, 6.7 ± 1.0 Hz, n = 11; STIM2 shRNA#1, 15.0 ± 0.6 pA, 2.9 ± 0.3 Hz, n = 9; STIM2 shRNA#1 + YFP-STIM2, 24.7 ± 1.6 pA, 6.4 ± 0.5 Hz, n = 11; **p < 0.01, ANOVA). (D) Cumulative distribution of mEPSC amplitude. (E) Synaptic density measured in control and STIM2-silenced hippocampal neurons (DIV 21) by costaining with presynaptic (VAMP2) and postsynaptic (Homer1) markers. Scale bar, 5 μm. Overlapping VAMP2/Homer1 puncta are detected and scored (F) by a MATLAB script (see Materials and Methods). Synaptic density per 10 μm: scr. shRNA, 6.1 ± 0.27, n = 15; STIM2 shRNA #1, 2.4 ± 0.3, n = 17; p < 0.01, t test.